LOCUS       sequence-418768-        4129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid expressing fusion construct of Gal4 DBD and VP16 AD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4129)
  AUTHORS   Budzynski MA, Puustinen MC, Joutsen J, Sistonen L
  TITLE     Uncoupling Stress-Inducible Phosphorylation of Heat Shock Factor 1 
            from Its Activation.
  JOURNAL   Mol Cell Biol. 2015 Jul;35(14):2530-40. doi: 10.1128/MCB.00816-14. 
            Epub 2015 May 11.
  PUBMED    25963659
REFERENCE   2  (bases 1 to 4129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/MCB.00816-14"; journalName: "Mol Cell Biol"; date: 
            "2015-07"; volume: "35"; issue: "14"; pages: "2530-40"
COMMENT     Sequence Label: Gal4-VP16
FEATURES             Location/Qualifiers
     source          1..4129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    343..477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(380..399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     434..453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(536..555)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(820..837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(991..1579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1071..1090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1750..2541,2542..2610))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2373..2392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2611..2715)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2770..2790)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             3112..3552
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3574..3807
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(3959..3978)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4003..4051
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
       61 acgtcggggc ggcaggccct gccatagcca ctggccccgt gggttaggga cggggtcccc
      121 catggggaat ggtttatggt tcgtgggggt tattattttg ggcgttgcgt ggggtcaggt
      181 ccacgaccca agcttgtcga cggtaccccg gggaattcga gctctagata agtaatgatc
      241 ataatcagcc atatcacatc tgtagaggtt ttacttgctt taaaaaacct cccacacctc
      301 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
      361 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
      421 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcctc
      481 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
      541 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
      601 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
      661 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
      721 ctaatgcagg agtcgcataa gggagagcgt cgatatgcgg tgtgaaatac cgcacagatg
      781 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
      841 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      901 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     1441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2041 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     2101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2341 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     2401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaccttctga ggcggaaaga
     2761 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2821 gaagtatgca aagcatgcat ctcaattagt cagcaaggaa agtccccagg ctccccagca
     2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2941 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3001 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3061 tgaagaggct tttttggagg agatctaagc ttgaagcaag cctcctgaaa gatgaagcta
     3121 ctgtcttcta tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa
     3181 gaaaaaccga agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa
     3241 accaaaaggt ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga
     3301 ctggaacagc tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg
     3361 gattctttac aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat
     3421 aaagatgccg tcacagatag attggcttca gtggagactg atatgcctct aacattgaga
     3481 cagcatagaa taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag
     3541 ttgactgtat cgccggaatt cccggggatc tgggcccccc cgaccgatgt cagcctgggg
     3601 gacgagctcc acttagacgg cgaggacgtg gcgatggcgc atgccgacgc gctagacgat
     3661 ttcgatctgg acatgttggg ggacggggat tccccgggtc cgggatttac cccccacgac
     3721 tccgccccct acggcgctct ggatatggcc gacttcgagt ttgagcagat gtttaccgat
     3781 gcccttggaa ttgacgagta cggtgggtag ggggcgcgac cggacccgca tcccccgtct
     3841 gggttttccc ctcccgtccc cggttcgtat ccacaataaa cacgagcaca tacattacaa
     3901 aaacctgcgg ttgtcgtctg attatttggt ggatccgggg gagatggggg aggctaactg
     3961 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4021 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4081 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcg
//
